Development of a Laboratory Transmission X-ray Microscope for Rapid 3D Cellular Tomography using Zernike Phase Contrast with 2.7 keV

使用 2.7 keV 泽尼克相差技术开发用于快速 3D 细胞断层扫描的实验室透射 X 射线显微镜

• 批准号: 10011500
• 负责人: David Vine
• 金额: $ 85.77万
• 依托单位: SIGRAY, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-15 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10011500
• 关键词: 3-Dimensional; Algae; Anodes; Area; Attenuated; Bacteria; Biological; Biology; Caliber; Carbon; Cell Nucleus; Cell physiology; Cells; Cellular Structures; Complement; Cryopreserved Cell; DNA; Development; Diagnostic radiologic examination; Diamond; Disease; Electrons; Fluorescence Microscopy; Hydration status; Image; Imaging Techniques; Laboratories; Legal patent; Light; Link; Mammalian Cell; Measures; Medical; Methods; Microscope; Microscopy; Molecular; Morphologic artifacts; Mutation; Operating System; Optics; Organelles; Oxygen; Performance; Phase; Photons; Proxy; RNA; Research; Resolution; Roentgen Rays; Science; Small Business Innovation Research Grant; Solid; Source; Stains; Structure; Synchrotrons; System; Systems Development; Techniques; Technology; Three-Dimensional Image; Three-Dimensional Imaging; Time; Water; X ray microscopy; Yeasts; absorption; base; cell water; cellular development; cellular imaging; contrast imaging; design; detector; high resolution imaging; imaging capabilities; imaging modality; improved; innovation; operation; preservation; prototype; screening; statistics; tomography; transmission process

Project Summary/Abstract X-ray microscopy has newly developed into an important new cellular imaging technique for visualizing and measuring intact cells in three dimensions. The development of these systems, enabled by advances in x-ray optic and detector technology, has been primarily driven at synchrotron light source facilities. The first laboratory x-ray microscopes for cellular imaging were introduced by the proposal’s co-PI in 2009 and utilized the absorption contrast of “water window” (285-540eV) x-rays for imaging. The system enabled unstained cellular tomography for the first time in the laboratory. However, the system operational energy is too low for most mammalian cells, which are larger in diameter than eukaryotic (yeast, bacteria, etc.) cells that the water window enables. We propose to develop a new 2.7 keV x-ray laboratory cellular microscope to provide fast (~30 minute) and high contrast complete 3D imaging of unstained cells of up to 80um in diameter and their organelles at 30nm resolution. The system operates using Zernike phase contrast at 2.7 keV energy x-rays, which provides higher contrast than even absorption contrast of water window x-rays. Additionally, the system will enable several major advantages over water window x-ray microscopy, including: much larger cell imaging (80µm vs. 10µm), larger depth- of-field for higher 3D resolution, and practical benefits (more stable x-ray source and larger working distance to incorporate correlative techniques). The microscope uses the company’s patented high brightness x-ray source and proprietary x-ray optic technology. The proposed Phase II 24-month project is to develop a complete prototype 2.7 keV Zernike phase contrast system for cellular imaging and to experimentally verify its performance.

项目摘要/摘要 X射线显微镜已新发展为重要的新蜂窝成像技术 在三个维度中可视化和测量完整的细胞。 由X射线光学和检测器技术的进步启用，主要驱动 同步子源设备。 提案的Co-Pi于2009年引入，并利用了“水”的吸收对比度 窗口“（285-540EV）用于成像的X射线。 但是，在实验室中，系统的运营能量太低了 哺乳动物细胞的直径比真核（酵母，细菌等）大的哺乳动物细胞大 水窗可以启用。 我们建议开发一个新的2.7 KEV X射线实验室细胞显微镜以提供快速（〜30 分钟）和高对比度完整的未染色细胞的3D成像，直径为80UM 及其细胞器以30nm的分辨率使用。 2.7 KEV Energy X射线，它提供的对比度比水的吸收对比度更高 窗户X射线。 窗户X射线显微镜，包含：更大的细胞成像（80µm vs. 10µm），深度较大 用于高3D分辨率的现场和实际好处（更稳定的X射线源和更大的 合并相关技术的工作距离）。 获得专利的高亮度X射线源和专有X射线光学技术。 支撑II期24个月的项目是开发一个Compesse Prototype 2.7 Kev Zernike 用于细胞成像和实验性验证其性能的相对比度系统。